Chemosphere 81 (2010) 577–583

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Chemosphere
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Assessing amendment and fertilizing properties of digestates from anaerobic

digestion through a comparative study with digested sludge and compost
Fulvia Tambone ⇑, Barbara Scaglia, Giuliana D’Imporzano, Andrea Schievano, Valentina Orzi, Silvia Salati,
Fabrizio Adani ⇑⇑
Gruppo RICICLA, Dipartimento di Produzione Vegetale, Università degli Studi di Milano, Via Celoria 2, 20133 Milano, Italy

a r t i c l e i n f o a b s t r a c t

Article history: Digestate, with biogas represents the final products of anaerobic digestion (AD). The methane-rich biogas
Received 22 April 2010 is used to produce electricity and heat, whereas the digestate could be valorized in agriculture. Contrarily
Received in revised form 13 August 2010 to well-recognized biomasses such as digested sludge and compost, the properties of the digestate are
Accepted 17 August 2010
not well known and its agricultural use remains unexplored.
Available online 9 September 2010
In this work, a first attempt to study the agronomic properties of digestates was performed by compar-
ing the chemical, spectroscopic, and biological characteristics of digestates with those of compost and
Keywords:
digested sludge, used as reference organic matrices. A total of 23 organic matrices were studied, which
Amendment properties
Anaerobic digestion
include eight ingestates and relative digestates, three composts, and four digested sludges.
Digestate The analytical data obtained was analyzed using principal component analysis to better show in detail
Fertilizer properties similarities or differences between the organic matrices studied.
CP MAS 13C NMR The results showed that digestates differed from ingestates and also from compost, although the start-
Principal component analysis ing organic mix influenced the digestate final characteristics. With respect to amendment properties, it
seems that biological parameters, more than chemical characteristics, were more important in describing
these features. In this way, amendment properties could be ranked as follows: compost ffi dige-
state > digested sludge  ingestate.
As to fertilizer properties, AD allowed getting a final product (digestate) with very good fertilizing prop-
erties because of the high nutrient content (N, P, K) in available form. In this way, the digestate appears to
be a very good candidate to replace inorganic fertilizers, also contributing, to the short-term soil organic
matter turnover.
Ó 2010 Elsevier Ltd. All rights reserved.

1. Introduction efficient and appropriate use in agriculture of organic fertilizers

The inputs of organic matter (OM) in the soil play a central role
in the productivity of arable land by providing nutrients, through
decomposition, and by maintaining soil fertility through OM turn-
over (Palm et al., 2001). The benefits of a balanced fertilization,
using organic amendment (e.g., crop residues, manure, compost)
in maintaining soil OM level in soils, have been increasingly
emphasized by researchers (Miller and Wali, 1995). At the same
time in the last decades, the use of organic wastes to produce or-
ganic fertilizers, with the aim of reducing dumping in landfill sites,
has grown a lot. Recycling of organic waste materials helps main-
tain soil nutrient levels, stimulating various aspects of soil fertility
(Iakimenko et al., 1996; Tambone et al., 2007). However, the
⇑ Corresponding author. Tel.: +39 02 50316547.
⇑⇑ Corresponding author. Tel.: +39 02 50316545.
E-mail addresses: fulvia.tambone@unimi.it (F. Tambone), fabrizio.adani@uni-
mi.it (F. Adani).
coming from organic wastes requires more in-depth knowledge
both in terms of quality and fertilizer value (Rowell et al., 2001).
Only in this way will it be possible to support crop production
and protect the environment, saving the soil resource (Mamo
et al., 1999). Recently, in European countries, among the biological
processes applied to treat organic wastes, interest in AD has in-
creased (Tani et al., 2006). AD is an anaerobic biological digestion
by which, in the absence of oxygen, OM is transformed into biogas,
which principally consists of methane (50–80 vol.%) and carbon
dioxide, the former used to produce energy and heat (Tani et al.,
2006). AD also produces a final biologically stable and partially hy-
gienic organic product, the digestate (Tambone et al., 2009).
AD has been reported to produce benefits by increasing the
agronomic value of the biomasses treated. For example, with re-
spect to animal slurry, AD allows the production of a digestate with
a higher proportion of mineral N and less decomposable OM
(Sørensen and Møller, 2009). This point is very important if one
considers that the N-fertilizing effect of slurry or similar material
is roughly equal to ammonium content and that the N leaching

0045-6535/$ - see front matter Ó 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.chemosphere.2010.08.034
578 F. Tambone et al. / Chemosphere 81 (2010) 577–583

from organic N mineralization depends on organic content (Søren- Table 1

sen and Møller, 2009). Composition of the biomasses studied.

No less important are the effects of the anaerobic digestion on Composition

the environment. AD-reduces emissions of greenhouse gas such w/w fresh matter
as methane and nitrous oxide (Sørensen and Møller, 2009). In addi- I1
tion AD contributes to global warming savings, not only from sub- I2 80% OFMSW + 20% pig slurry
stitution of fossil fuel by biogas but also from carbon storage in the I3

soil and inorganic fertilizer substitution (Møller et al., 2009). I4 48% Pig slurry + 24% milk serum + 14% cow slurry + 10% maize
Therefore, a strong contributing factor to the success of AD comes I5 silage + 4% rice residues

from the valuation of the digestate in agriculture (Møller et al., I6

2009). Nevertheless, in contrast to well-recognized biomasses such I7 65% Pig slurry + 20% blood industry residues + 15% maize silage
I8
as digested sludge (DS) and compost, the digestate is not well
known and its agricultural use remains an unexplored field in re- C1 Green compost: 100% ligno-cellulosic residues

search. Digestates, in fact, have not undergone scientific examina- v/v fresh matter
tion that will reveal their configuration in agronomic terms. C2
Mixed composts: ligno-cellulosic residues plus OFMSW in a 1:2 ratio
C3
Whether digestates could act more as fertilizer or amendment or
both is still not clear. Having amendment properties implies that DS1
DS2 Anaerobically treated and dried digested slurries from municipal
the organic matrices contain organic fractions that can contribute DS3 wastewater
to SOM turnover, as this latter process influences biological, chem- DS4
ical, and physical soil characteristics (Lal, 2001). On the other hand,
fertilizer properties refer to the amount of nutrient elements con-
tained in the organic matrices and their chemical forms (Rowell
et al., 2001).
three full-scale plants: one produces green compost from ligno-
DSs, for example, have been extensively studied in the past (Ia-
cellulosic residues, and two produce mixed composts from ligno-
kimenko et al., 1996) and, because of their high N and P content
cellulosic residues plus the organic fraction of municipal solid
and low C:N, they represent an organic matrix used more for its
waste (OFMSW) obtained by separate collection, in a 1:2 (v/v) fresh
fertilizer properties, with amendment effects being visible only at
matter ratio.
very high dosage (Iakimenko et al., 1996).
On the other hand, compost is used more for its amending prop-
2.2. Chemical analyses
erties because of its low nutrient content and high OM quality
(Giusquiani et al., 1995; Tambone et al., 2007). Research has shown
All samples collected, about 5 kg each, were brought to the lab-
that compost may improve the chemical and physical properties of
oratory, dried for 24 h at 40 °C and then for another 24 h at 105 °C
the soil by increasing nutrient and water capacity, OM content, pH,
(APHA, 1992), shredded in a blender and passed through a 1-mm
and cation exchange capacity (Giusquiani et al., 1995; Leifeld et al.,
mesh. The samples were successively subjected to biological,
2002) and that the effects of compost depend on the dose used
chemical, and spectroscopic analyses. NH3 and TKN were detected
(Tambone et al., 2007).
on fresh samples.
Both amendment and fertilizer properties of organic matrices
Fresh matter (FM), total solids (TS), volatile solids (VS), and total
can be revealed by long-term, full-field studies (Adani and Tam-
organic carbon (TOC) were determined following standard proce-
bone, 2005). Nevertheless, this approach is very expensive and
dures (APHA, 1992). Ammonia and TKN were determined using
time-consuming. In this work, a first attempt to investigate the
the analytical method for wastewater sludges (IRSA CNR, 1994).
agronomic properties of digestates was performed by studying
Organic N was calculated as the difference between TKN and
their chemical, spectroscopic and biological characteristics in com-
NH3. Total P and K contents were determined by inductively cou-
parison with those of compost and DS, which were used in this
pled plasma mass spectrometry (Varian, Fort Collins, USA). Stan-
work as reference organic matrices. Results obtained in this work
dard samples (National Institute of Standards and Technology,
could be of help in the preparation of a guidelines for the digestate
Gaithersburg, MD, USA) and blanks were run with all samples to
use in agriculture. By now, there is not a EU regulation for the use
ensure precision in the analyses. P and K detection was preceded
of digestate and only local and on volunteer basis guidelines have
by acid digestion (EPA, 1998) of the biomass samples. All analyses
been proposed. For example the British Standards Institution (BSI)
were performed in triplicate.
(2010), published a volume in which indications for a correct use of
digestate are reported. On the other hand, the European Commis-
13
sion is considering proposing minimum standards and guidelines 2.3. CP MAS C NMR analysis
for use of digestate in agriculture via the revision of the Sewage
Sludge Directive. The solid-state Cross Polarization Magic Angle Spinning 13C Nu-
clear Magnetic Resonance (CP MAS 13C NMR) spectra of the sam-
ples were acquired at 10 kHz on a Bruker AMX 600 spectrometer
2. Material and methods (Bruker BioSpin GmbH, Rheinstetten) using a 4-mm CP MAS probe.
The pulse repetition rate was set at 0.5 s, the contact time at 1 ms,
2.1. Organic matrices studied and the number of scans was 3200. The chemical shift scale of CP
MAS 13C NMR spectra were referred to tetramethylsilane
The biomasses studied (Table 1) were as follows: eight inge- (d = 0 ppm). Spectra were elaborated using TOPSPIN 1.3 software
states (I) and the corresponding eight digestates (D) (Pognani (Bruker BioSpin GmbH, Rheinstetten, Germany).
et al., 2009), four DS, and three composts (C), for a total of 23 sam-
ples. The ingestates and digestates were sampled in 2008 at three 2.4. Biological stability determination
different AD plants that produce biogas.
Anaerobically treated and dried DSs came from two municipal Biological stability was determined by a long-term degradation
wastewater treatment plants. The composts were collected from test (60 d) using the anaerobic potential biogas production test
 F. Tambone et al. / Chemosphere 81 (2010) 577–583 579

(ABP test) (Schievano et al., 2008). Anaerobic test has been re-
ported to describe OM degradability well (Schievano et al., 2008).
In brief, in a 100 mL serum bottle, 0.62 g of dried sample
(Ø < 1 mm) was added to 37.5 mL of inoculum and 22 mL of deion-
ized water. The batch tests were carried out with 60 mL samples
(about 35 g kg1 TS) and 40 mL of headspace. The fresh feedstock
and inoculum percentages of TS were respectively 35% and 65%.
Control blanks were prepared using 60 mL of inoculum.
All batches were sealed with Teflon hermetic caps, flushed with
an N2 atmosphere, and incubated at 37 ± 1 °C, until no further bio-
gas production was detected (normally around 60 d).
work, principal component analysis (PCA) was used to compare
biomasses qualitatively. PCA has been conducted on the log-trans-
formed data (Pichler et al., 2001). The determinant value, Bartlett’s
test of sphericity, and the Kaiser–Meyer–Olkin test of sampling
adequacy were initially performed on the data to evaluate the
appropriateness of conducting PCA. The size of each component
was identified by the eigenvalue: the earlier (and more significant)
the component, the larger its size. Only the PCs with eigenvalues
>1 were retained (Kaiser, 1960; Brejda et al., 2000). All statistical
analyses were carried out using SPSS statistical software (vers.
16) (SPSS, Chicago, IL).

2.5. Statistical approach 3. Results

Chemical analyses were performed in triplicate on each bio-
mass sample taken from 5 kg representative mass. Since chemical
analyses were performed on three analytical samples withdrawn
from the 5 kg composite bulk sample, standard deviation values
calculated from the data for three replications were estimates of
the variability caused by both the analytical method and the bulk
sample homogeneity. Average and standard deviation values from
chemical analyses were calculated according to standard proce-
dures and the results analyzed by ANOVA. Tukey’s test was used
to compare mean values and to assess the significance of the differ-
ences between mean values.
Biomass similarity was studied by using chemical, spectro-
scopic, and biological parameters detected for each biomass. Thus,
the large amount of information required a reduction in dimen-
sionality (generally two dimensions). As a consequence, the analyt-
ical data obtained were elaborated by multivariate analysis. In this
3.1. Chemical characteristics
The chemical characteristics for ingestates, digestates, DSs and
composts are reported in Table 2. Results, reported as average of
sample data composing each organic matrix, showed that the inge-
states had the highest TOC and VS contents. These data support the
fact that ingestates did not undergo any kind of biological process.
In contrast, anaerobic (digestates) or aerobic (composts) or the
combination of the two biological treatments (digestate sludge)
were responsible for the partial mineralization of OM, thus the
lower TOC and VS contents of digestates, DSs, and composts (Adani
et al., 2006; Tambone et al., 2009). Composts, in particular, showed
the lowest TOC values, indicating a great degree of mineralization
of OM (Lee, 1992), although both TOC and VS were affected also by
the composition of the starting organic matrices used to produce
the compost.

Table 2
Chemical and biological characteristics of the samples studied in this work.

TS VS (g kg1 TS) TOC TKN NH3–N NH3–N Org-Na C:Nb P2O5 K2O ABP (normal
(g kg1 FM) (g kg1 TS) (g kg1 TS) (g kg1 TS) (% TKN) (g kg1 TS) (g kg1 TS) (g kg1 TS) L kg1 VS)
I1 110 ± 10 823 ± 8 451 ± 7 55 ± 1 17 ± 1 32 37 12 15 ± 1 27 ± 2 649 ± 50
I2 112 ± 4 873 ± 9 482 ± 2 32 ± 1 8±1 26 24 20 10 ± 1 16 ± 3 732 ± 58
I3 123 ± 6 920 ± 18 510 ± 5 39 ± 2 19 ± 2 48 20 25 12 ± 1 18 ± 1 714 ± 21
I4 119 ± 7 883 ± 5 473 ± 2 28 ± 1 12 ± 2 43 16 29 27 ± 2 13 ± 1 631 ± 4
I5 146 ± 7 882 ± 3 492 ± 4 24 ± 1 11 ± 2 45 13 37 27 ± 0 19 ± 2 596 ± 26
I6 144 ± 8 849 ± 2 492 ± 9 27 ± 2 16 ± 1 59 11 46 35 ± 1 39 ± 1 375 ± 14
I7 115 ± 3 822 ± 11 477 ± 6 44 ± 2 23 ± 1 51 21 22 27 ± 1 55 ± 1 449 ± 12
I8 127 ± 9 852 ± 18 494 ± 5 44 ± 1 18 ± 1 41 26 19 25 ± 0 49 ± 1 396 ± 13
Mean n=8 124 ± 14bc 863 ± 33c 484 ± 18c 37 ± 11ab 15 ± 5b 43 ± 11c 21 ± 8b 26 ± 11b 22 ± 9a 30 ± 16c
568 ± 142b
D1 45 ± 3 707 ± 1 397 ± 9 135 ± 2 87 ± 3 63 48 8 17 ± 2 59 ± 1 399 ± 27
D2 38 ± 6 685 ± 17 384 ± 11 145 ± 2 97 ± 1 68 48 8 21 ± 1 48 ± 1 396 ± 56
D3 31 ± 12 681 ± 19 378 ± 10 151 ± 1 98 ± 1 65 53 7 35 ± 2 58 ± 2 372 ± 79
D4 32 ± 8 696 ± 28 367 ± 8 103 ± 1 63 ± 2 61 41 9 52 ± 2 29 ± 2 240 ± 48
D5 44 ± 3 702 ± 30 383 ± 2 83 ± 2 45 ± 2 54 38 10 53 ± 2 36 ± 2 281 ± 15
D6 53 ± 1 667 ± 1 387 ± 8 85 ± 2 51 ± 2 61 34 11 58 ± 1 82 ± 0 211 ± 6
D7 63 ± 1 722 ± 1 418 ± 7 86 ± 3 54 ± 1 62 32 13 53 ± 1 67 ± 2 184 ± 15
D8 60 ± 0 735 ± 3 421 ± 9 92 ± 1 61 ± 2 67 31 14 50 ± 1 55 ± 2 193 ± 65
Mean n=8 46 ± 12a 670 ± 22b 392 ± 19b 110 ± 29c 69 ± 21c 63 ± 4d 41 ± 8c 10 ± 2a 43 ± 16ab 54 ± 17d
284 ± 92a
C1 769 ± 4 362 ± 5 184 ± 8 12 ± 1 1±0 10 11 17 6±1 9±2 517 ± 58
C2 731 ± 7 679 ± 11 284 ± 5 19 ± 0 2±0 10 17 16 29 ± 1 21 ± 2 323 ± 26
C3 711 ± 11 415 ± 7 200 ± 3 17 ± 1 2±0 10 15 13 31 ± 1 16 ± 1 472 ± 58
Mean n=3 737 ± 30d 485.5 ± 170.0a
222 ± 54a 16 ± 4a 2 ± 0a 10 ± 0a 14 ± 3a 15 ± 3ab 22 ± 14a 15 ± 6b 437 ± 102ab
DS1 176 ± 5 765 ± 12 412 ± 10 73 ± 1 14 ± 0 19 59 7 52 ± 1 5±0 438 ± 15
DS2 145 ± 3 743 ± 9 363 ± 9 55 ± 0 10 ± 2 19 45 8 50 ± 1 7±1 385 ± 18
DS3 185 ± 6 682 ± 14 357 ± 11 50 ± 2 9±2 18 41 9 40 ± 1 4±0 359 ± 58
DS4 131 ± 11 660 ± 7 340 ± 3 50 ± 2 9±1 19 41 8 42 ± 2 6±0 394 ± 29
Mean n=4 159 ± 25c 712 ± 50b 368 ± 31b 57 ± 11b 11 ± 2b 18 ± 0b 47 ± 9c 8 ± 0a 46 ± 6ab 5 ± 1a
394 ± 33ab
a
Calculated as TKN–NH3–N.
b
Calculated as TOC/org-N.
c
Means followed in the same column by the same letter are not statistically different (p < 0.05) according to Tukey’s test.
580 F. Tambone et al. / Chemosphere 81 (2010) 577–583

Total N content differed among the biomasses studied (Table 2)
and followed this order: digestates > DSs > ingestates > composts.
Data in Table 2 reflected the typical N contents of these biomasses,
which depended on both the composition of the starting organic
matrix and the biological processes that the matrices underwent.
The higher N content of the digestate with respect to the ingestate
was a consequence of the N concentration effect, because of carbon
degradation to CO2 and CH4 and N preservation during AD (Tam-
bone et al., 2009). More interesting were the data on the reparti-
tion of TKN into ammonia and organic N (Table 2). During AD,
OM mineralization led to the organic transformation of the organic
N into ammonia, with the consequent increase in NH3 content in
the digestate. The low TKN content and, above all, the low NH3
content measured in the composts were expected, as the aerobic
treatment determined ammonia losses by its stripping during
composting if pH is high (Cadena et al., 2009). On the other hand,
the low TKN and ammonia of the DS was due to the drying process
that occurred at the end of the wastewater treatment, allowing
ammonia to be volatilized.
Among the mechanisms that regulate OM mineralization and
conservation in the soil, a determinant role is assumed by the
C:N ratio (Barrett and Burke, 2000). The C:N values (Table 2) were
calculated, taking into account only the organic N fraction as the
NH3 fraction did not contribute to the medium and long-term N
turnover in the soil; this fraction was rapidly lost because of plant
uptake, ammonia volatilization in the air, or nitrate leaching into
the soil (Huang and Chen, 2009). The C:N of digestates and DSs
were similar and much lower than those of composts (Table 2).
Concerning the P (P2O5) content of organic matrices, digestates
and DSs showed the same concentrations of this element (Table 2),
twice that of the compost, that was similar to that of the ingestates.
Digestates also had the greatest concentration of K (K2O) of all the
other organic matrices (Table 2).
3.2. CP MAS 13
C NMR analysis
CP MAS 13C NMR provides qualitative and quantitative informa-
tion on the composition of the ingestates, digestates, DSs and com-
posts. It identifies the main C type that comprises OM (Conte et al.,
1997). Here, this technique was applied to verify the changes in
macromolecular composition that occur in the ingestates through
AD treatment and to measure the differences and/or similarities
between digestates, composts, and DSs.
Five types of carbon can be distinguished in the NMR spectrum
(Table 3): (i) short chain aliphatic carbon, e.g., volatile fatty acids
(VFAs) and steroid-like molecules (Réveillé et al., 2003) (0–
28 ppm); (ii) long chain aliphatic carbon (e.g., plant aliphatic bio-
polymers) (Pereira et al., 2005) and proteins (Dignac et al., 2000)
(28–47 ppm); (iii) O-alkyl carbon (e.g., polysaccharides) (Kogel-
Knaber, 2002) (47–113 ppm); (iv) aromatic carbon (e.g., lignin)
(Ussiri and Johnson, 2003) (113–160 ppm); and (v) carbonyl car-
bon in aliphatic esters, carboxyl groups, and amide carboxyl
(160–210 ppm).
The ingestates’ NMR (Table 3) were well-characterized by the
presence of the 0–47 ppm area, which indicated the presence of
both long chain linear structures (e.g., suberin, cutin, waxes, fatty
acid) (peaks at 30–33 ppm) derived from animal slurry, municipal
solid waste, and energy crops (Dignac et al., 2000; Pichler et al.,
2001) and of branched or short chains (peaks at 24–25 ppm), such
as steroid-like molecules, most likely derived from animal slurry
preserved during the biological treatments (Réveillé et al., 2003).
What was interesting in the spectra of ingestates was the presence

Table 3
13
CP MAS C NMR integrated area of the samples studied in this work.

C-type
Aliphatic-C bonded to other aliphatic chain or to H Total aliphatic-C O–CH3 or N-alkyl Aromatic-C phenol or Carboxyl-C keto-C
O-alkyl-C di-O-alkyl-C phenyl ether-C
Short chain Long chain
Band d range (ppm)
0–28 28–47 0–47 47–113 113–160 160–210
I1 12.7 9.3 21.9 60.7 6.6 10.7
I2 13.4 16.4 29.8 56.9 5.1 8.1
I3 17 8.9 26 59.6 3.7 10.7
I4 11.1 7.7 18.9 67.2 6.8 7.2
I5 11.2 7.1 18.4 66.6 7.1 7.8
I6 9.1 10 19.2 67.8 7.7 5.3
I7 10.9 8.9 19.7 64.8 8.7 6.8
I8 9.2 11.4 20.6 64.9 7.6 6.8
Mean n = 8 11.8 ± 2.6 10 ± 3 21.8 ± 4aa 63.6 ± 4c 6.7 ± 1.6a 7.9 ± 1.9a
D1 19.5 23.3 42.8 37.5 8.5 11.2
D2 23.6 23.1 46.7 32.7 7.5 13.1
D3 21.1 22.9 44 36 7.7 12.2
D4 11.9 13.7 25.6 59.4 7.9 7.1
D5 13.4 14.8 28.1 55.3 8.1 8.4
D6 13.2 17.1 30.3 54.4 8 7.2
D7 18.2 20.3 38.5 44.1 7.7 9.6
D8 12.2 16.6 28.8 55.2 8.6 7.4
Mean n = 8 16.6 ± 4.5 19 ± 4 35.6 ± 8.3b 46.8 ± 10.5ab 8 ± 0a 9.5 ± 2.4ab
C1 nd nd 25 53.4 12.8 8.8
C2 nd nd 22.3 56 13.14 8.5
C3 nd nd 17.9 56.9 15.72 9.5
Mean n = 3 nd nd 21.7 ± 3.6a 55.4 ± 1.8bc 13.9 ± 1.6b 8.9 ± 0.5ab
DS1 17.5 20.3 37.8 42.8 7 12.3
DS2 18.5 18.6 37.1 44.8 6.3 11.8
DS3 17.1 18.7 35.8 44.7 7.2 12.3
DS4 17.9 20.4 38.3 42.8 7 11.8
Mean n = 4 17.7 ± 0.6 19.5 ± 1 37.3 ± 1.1b 43.8 ± 1.1a 6.9 ± 0.4a 12.1 ± 0.3b
a
Means followed in the same column by the same letter are not statistically different (p < 0.05) according to Tukey’s test.
 F. Tambone et al. / Chemosphere 81 (2010) 577–583
of the peaks at 21 ppm, indicating the presence in the biomass of
VFAs, which were probably formed during biomass storage under
semi- or anaerobic condition. This peak disappeared in the dige-
state, indicating VFA degradation during AD (Tambone et al.,
2009). The ingestates’ spectra were also dominated by the pres-
ence of carbohydrate molecules (e.g., hemicelluloses, celluloses,
and simple sugars) (see 47–113 ppm area) (Table 3). The strong
reduction of this area in the digestates suggests that AD proceeded,
above all, through the degradation of the carbohydrate fraction
(Tambone et al., 2009).
As a consequence of the degradation of the labile organic mol-
ecules during AD (i.e., VFAs and O-alkyl-C), the digestates’ spectra
(Table 3) showed an increase in NMR signals in the 0–47 ppm and
130–160 ppm regions. This means that the more recalcitrant frac-
tions (plant biopolymers, steroid-like molecules, and lignin) con-
tained in the starting organic mixtures were concentrated
(Tambone et al., 2009).
DSs showed a macromolecular composition similar to that of
digestates. The presence of recalcitrant biogenic molecules such
as steroids from human feces (Réveillé et al., 2003) and those of
non-biogenic origin in the domestic wastewaters (Schnaak et al.,
Fig. 1. PCA plots for amendment properties (a) and fertilizer properties (b).
581
1997) were probably responsible for the high alkyl-C content in
DSs (see 0–47 ppm region) (Table 3).
The composts (Table 3) showed macromolecular compositions
different from other organic matrices studied, i.e., higher O-alkyl-
C and aryl-C contents and lower alkyl-C content, than those of
other organic matrices (Table 3). Lignocellulosic materials used in
the starting mixer as bulking agents determined higher lignin-like
and O-alkyl-Cs contents in the final products (composts) (Dinel
et al., 1998; Ussiri and Johnson, 2003) than those in both digestates
and ingestates.
3.3. Biological stability
The ABP production of the organic matrices studied (Table 2)
showed that the ingestates were the less degraded biomasses
(Schievano et al., 2008; Tambone et al., 2009). On the contrary, bio-
logically treated biomasses such as composts, DSs, and digestates
showed lower biogas production than ingestate (Table 2) Dige-
states showed ABP values lower than both compost and above all
DS, although average values were not statistically different (Table
2), and similar to those reported in a previous study (Schievano
et al., 2008). In particular digestate data indicate a high degree of
biological stability (Tambone et al., 2009).
3.4. PCA
In this study, PCA was applied to two different data sets with
the aim of studying both amendment (PCAM) and fertilizer proper-
ties (PCF) of the biomasses.
Data sets relative to OM contents (VS and TOC), biological sta-
bility (ABP test) (Table 2), and macromolecular composition (CP
MAS 13C NMR) (Table 3) were considered useful for measuring
the amendment properties. The second data set, related to the fer-
tilizing properties, included TKN, NH3, org-N, C:N, total P2O5, and
total K2O contents (Table 2).
The determinant values (0.00004 and 0.0001 for PCAM and PCF,
respectively) and the results of Bartlett’s test of sphericity (signif-
icance of 0.0001 for both PCAM and PCF) confirmed the appropriate
level of correlation to perform PCA (Anderson et al., 2004). In addi-
tion, the Kaiser–Meyer–Olkin test of sampling adequacy measure,
which was 0.5 for both PCAM and PCF, confirmed the sampling ade-
quacy of the data set used (Anderson et al., 2004).
The PCA of amendment properties gave two principal compo-
nents that are able to explain 79% of the total variability of the sys-
tem (Fig. 1). The use of the data set relative to fertilizing properties
and the successive application of PCA allowed the extraction of two
principal components (PCF1 and PCF2) that explained 82% of the
system’s total variability (Fig. 1).
4. Discussion
Anaerobic processes are conducted with the aim of producing
biogas. In this way and in order to maximize biogas production,
easily degradable biomasses such as animal slurries, OFMSW,
energy crops, and agro-industrial residues are used alone or in
mixture. AD, if correctly performed, proceeds by the high degrada-
tion of OM (Pognani et al., 2009). Consequently, starting organic
matrices are transformed during AD. OM transformation involves
the preservation of recalcitrant molecules that are concentrated
(Tambone et al., 2009). CP MAS 13C NMR gave evidence of that,
indicating that both lignin-like material and complex lipids and
steroids became concentrated during AD. These molecules have
been reported to be humus precursors (Lorenz et al., 2007), playing
an important role in the short-term SOM turnover. On the other
hand, because of mineralization of OM and of the anaerobic
582 F. Tambone et al. / Chemosphere 81 (2010) 577–583

condition in which AD is performed, the preservation and concen- the compost centroid (Fig. 1), one can surmise for each matrix
tration of inorganic nutrients, such as P and K and N, occurred group a sort of ‘‘compost similarity”, which is as follows: C1, C2,
(Pognani et al., 2009). In addition, organic N is transformed into C3 > D4, D5, D6, D8  I4, I5, I6, I7, I8 ffi D1, D2, D3, D7, DS3, DS2,
ammonia (Pognani et al., 2009). DS1, DS4 > I1, I2, I3.
The data of this study confirmed that digestates are very differ- On the basis of these results, it can be suggested that digestates
ent from ingestates and, both OM quality and fertilizing element show characteristics different (D4, D5, D6, D8 – D1, D2, D3, D7)
content of the digestates suggest its usefulness in agriculture from those of compost. As D1–D3 digestates were derived from
(Tambone et al., 2009). In this study, to know the digestate’s I1–I3 ingestates, which were different from other ingestates in
amendment and fertilizing properties, a comparison was made be- terms of materials composing the mix (Table 1) and chemical com-
tween digestates and well known organic matrices. Thus compost position (Tables 2 and 3), it can be concluded that the origin and
was used in the study as it is one of the most classical organic composition of the organic matrices influenced the digestate’s
amendments studied and DS was chosen because it has a high con- chemical characteristics. In particular, it seems that the presence
tent of nutritional elements such as N, P, and K. of the organic fraction of municipal solid wastes (Table 1) gave dig-
The results of this study reveal differences in the chemical com- estates characteristics much different from those of compost com-
position of OM and nutrients contained in the digestate and the pared with digestates obtained from a mix with animal slurry or
ingestate and, more importantly, differences with respect to the crop energy biomasses prevalent (Table 1). These data appear
DSs and composts. Considering that compost is an organic amend- strange as two of the three composts were obtained from a mix
ment and that DS is a nutrient fertilizer, the chemical, spectro- containing OFMSW (such as D1–D3), suggesting greater similarity.
scopic, and biological data of the samples studied were analyzed On the other hand, the three composts studied did not show big
using multivariate analysis (PCA) to detect similarities or differ- differences in composition among them, although C1 came from
ences among the four types of biomasses. lignocellulosic material and did not contain OFMSW. This fact indi-
The matrix of the components (Table 4), with respect to amend- cates that the lignocellulosic material present in the compost mix,
ment properties, shows that the PCAM1 (which explained 43% of to- more than OFMSW, contributed to the final compost composition
tal variability) was directly correlated to alkyl-C and carboxyl-C and that this material strongly affected compost compositions
contents and inversely correlated to the O-alkyl-C content, i.e., bio- resulting in a material completely different from the other organic
logical process-concentrated alkyl-C and carboxyl-C and degraded matrices (Fig. 1).
O-alkyl-C. In any case, digestate properties seemed to be very different
The second component (PCAM2) (which explained 36% of total from those of ingestates (see Euclidean distance between I and D
variability) was directly correlated to VS, TOC, and inversely corre- samples) (Fig. 1). It was interesting to note that, with respect to
lated to the content of aromatic-C, i.e., the more the matrices were the compost (Euclidean distances) some digestates (samples D1,
mineralized, the more lignin-like molecules were contained. D2, D3, D7) were similar to the ingestates (similar Euclidean dis-
The distribution of biomasses in the bi-dimensional space, de- tance). Nevertheless, the similarity was brought about by different
fined by the two PCPs, shows a clear distinction of the composts causes. Samples D1, D2, D3, and D7 were closer to the compost in
from the other biomasses (Fig. 1). This result can be attributed terms of biological stability (ABP) and OM content (VS and TOC
not only to the different biological processes (aerobic vs. anaero- contents) (Table 2), but they were far from it in terms of macromo-
bic) which the compost has undergone, but also to the presence lecular composition (alkyl-C and O-alkyl-C contents) (Table 3). In
of lignocellulosic materials used as bulk agent in the mixers stud- contrast, the ingestates were more similar to compost with regard
ied; this contributed greatly to the presence of aromatic-C in the to macromolecular composition but not with OM and ABP (Tables
final compost (Ussiri and Johnson, 2003). Digestates were posi- 2 and 3).
tioned in the PCA bi-plot some what close to the DS (samples D1, The ability of an organic matrix to contribute actively to the
D2, and D3) (Fig. 1) and partly intermediate between DS and com- SOM in a short period depends on the presence of recalcitrant mol-
posts (samples D4, D5, D6, and D8) (Fig. 1). Sample D7 was inter- ecules (Adani and Tambone, 2005). The degree of recalcitrance of
mediate between the two digestate groups. In Fig. 1, five organic organic biomass can be measured by detecting its biological degra-
matrix groups could be seen: C1–C3; D4, D5, D6, D8; D1, D2, D3, dability (biological stability) (Adani et al., 2004; Scaglia et al.,
D7, DS1, DS2, DS3, DS4; I1, I2, I3; and I4, I5, I6, I7, I8. The centroid 2010).
(Scaglia and Adani, 2008) for each organic matrix group was given From the results of this study, it appears that macromolecular
in Fig. 1. Taking into consideration the Euclidean distances from composition is not a suitable approach to test the recalcitrance/
biological stability of OM. For example, ingestates showed very
low biological stability but a macromolecular composition similar
Table 4 (e.g., alkyl-C content) to those of composts, which, contrastingly
Component loadings matrix of the PCAs for the amending properties (PCAM) and for showed low degradability. On the other hand, digestates D1, D2,
fertilizer properties (PCF) data sets. D3, and D7, although having a macromolecular composition differ-
Data sets PCAM PCF ent from those of composts, showed high biological stability simi-
lar to that of composts. Therefore, it could be concluded that
PCAM1 PCAM2 PCF1 PCF2
chemical composition, such as the one detected in this work, did
VS 0.15 0.96 – –
not greatly contribute to elucidating the digestate amendment
TOC 0.14 0.97 – –
ABP 0.10 0.12 – – properties by direct comparison with compost. Probably, biological
Total aliphatic-C 0.94 0.11 – – stability or biomass degradability is a more appropriate parameter.
O–CH3 or N-alkyl, O-alkyl C, di-O-alkyl C 0.96 0.13 – – In this work, ABP was chosen as a tool to detect biological stability.
Aromatic-C phenol or phenyl ether-C 0.21 0.88 – – On the basis of this parameter and taking into consideration
Carboxyl-C and keto-C 0.91 0.1 – –
TKN – – 0.83 0.53
the average ABP data reported in Table 2, the amendment proper-
NH3–N – – 0.18 0.94 ties could be ranked in the following manner: compost ffi
Org-N – – 0.97 0.07 digestate > DS  ingestate.
C/N – – 0.92 0.22 The matrix components (Table 4) relative to the PCA performed
P2O5 – – 0.57 0.19
to study fertilizing properties of the organic matrices showed that
K2O – – 0.02 0.91
PCF1 was directly correlated to TKN and organic N contents and in-
 F. Tambone et al. / Chemosphere 81 (2010) 577–583
versely correlated to C:N. The second component (PCF2) was di-
rectly correlated to the NH3, P2O5, and K2O contents.
The distribution of the biomasses in the bi-dimensional space,
defined by the two PCFs (Fig. 1), resulted in a good separation of
the four typologies of biomasses. In particular, taking into account
the PCA results and the data in Table 2, it can be concluded that all
digestate samples were characterized by the greatest TKN, NH3,
P2O5, and K2O contents and so they showed the highest nutrient
values. On the other hand, compost showed the opposite charac-
teristics as expected. The ingestate characteristics did not influence
the digestate properties.
Acknowledgements
This study was supported by Lombardy Region – National Pro-
grams for Biofuel (Probio Biogas Project – Regione Lombardia,
Italy) – and ARAL – Associazione Regionale Allevatori della Lom-
bardia contract N. Prot. USM 0032370.
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